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Glycoconjugate Journal

, Volume 32, Issue 3–4, pp 141–151 | Cite as

Identification of β1,3-galactosyltransferases responsible for biosynthesis of insect complex-type N-glycans containing a T-antigen unit in the honeybee

  • Tomomi Ichimiya
  • Megumi Maeda
  • Shou Sakamura
  • Masato Kanazawa
  • Shoko NishiharaEmail author
  • Yoshinobu KimuraEmail author
Original Article

Abstract

Honeybees (Apis mellifera) produce unique complex-type N-glycans bearing a Galβ1-3GalNAc (T-antigen) unit, and honeybee-specific N-glycans are linked to royal jelly glycoproteins. In this study, we identified two novel honeybee β1,3-galactosyltransferase (β1,3-GalT) genes responsible for biosynthesis of the T-antigen in insect N-glycans. The products of the two putative β1,3-GalT genes (β1,3-GalT1 and β1,3-GalT2), which were expressed in Sf21 insect cells, transferred galactose (Gal) residues to GalNAc2GlcNAc2Man3GlcNAc2-PA to form the Galβ1-3GalNAc unit, indicating that the identified genes were involved in biosynthesis of the β1-3 Gal-containing N-glycan. Therefore, using biochemistry and molecular biology techniques, we revealed a unique N-glycan biosynthesis mechanism in the cephalic region of honeybees, which has not previously been found in other animal or plant cells.

Keyword

Insect β1,3-galactosyltransferase Insect N-glycan N-glycan biosynthesis Thomsen-Friedenreich antigen Apis mellifera 

Abbreviations

β1,3-GalT

β1,3-galactosyltransferase

GalNAc α-pNph

p-nitrophenyl-N-acetyl-α-galactosaminide

MRJP

Major royal jelly protein

PA-

Pyridylamino

SF-HPLC

Size-fractionation HPLC

Man

d-mannose

Gal

d-galactose

GlcNAc

N-acetyl-d-glucosamine

GalNAc

N-acetyl-d-galactosamine

LacdiNAcM3

GalNAcβ1-4GlcNAcβ1-2Manα1-6(GalNAcβ1-4GlcNAcβ1-2Manα1-3) Manβ1-4GlcNAcβ1-4GlcNAc-PA

GalLacdiNAcM3

Galβ1-3GalNAcβ1-4GlcNAcβ1-2Manα1-6(GalNAcβ1-4GlcNAcβ1-2Manα1-3) Manβ1-4GlcNAcβ1-4GlcNAc-PA or GalNAcβ1-4GlcNAcβ1-2Manα1-6(Galβ1-3GalNAcβ1-4GlcNAcβ1-2Manα1-3) Manβ1-4GlcNAcβ1-4GlcNAc-PA

Gal2LacdiNAcM3

Galβ1-3GalNAcβ1-4GlcNAcβ1-2Manα1-6(Galβ1-3GalNAcβ1-4GlcNAcβ1-2Manα1-3) Manβ1-4GlcNAcβ1-4GlcNAc-PA

T-antigen

Thomsen-Friedenreich antigen (Galβ1-3GalNAc)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Tomomi Ichimiya
    • 1
  • Megumi Maeda
    • 2
    • 3
  • Shou Sakamura
    • 3
  • Masato Kanazawa
    • 3
  • Shoko Nishihara
    • 1
    Email author
  • Yoshinobu Kimura
    • 2
    • 3
    Email author
  1. 1.Department of Bioinformatics, Faculty of EngineeringSoka UniversityTokyoJapan
  2. 2.Department of Biofunctional Chemistry, Graduate School of Environmental and Life ScienceOkayama UniversityOkayamaJapan
  3. 3.Department of Biofunctional Chemistry, Graduate School of Natural Science and TechnologyOkayama UniversityOkayamaJapan

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